JP7611465B2 - Eluent-based fraction collector - Google Patents
Eluent-based fraction collector Download PDFInfo
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- JP7611465B2 JP7611465B2 JP2019187088A JP2019187088A JP7611465B2 JP 7611465 B2 JP7611465 B2 JP 7611465B2 JP 2019187088 A JP2019187088 A JP 2019187088A JP 2019187088 A JP2019187088 A JP 2019187088A JP 7611465 B2 JP7611465 B2 JP 7611465B2
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- 239000003480 eluent Substances 0.000 title description 15
- 238000000926 separation method Methods 0.000 claims description 48
- 239000011521 glass Substances 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 24
- 239000007788 liquid Substances 0.000 claims description 14
- 238000004587 chromatography analysis Methods 0.000 claims description 11
- 230000005484 gravity Effects 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 3
- 230000001737 promoting effect Effects 0.000 claims 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 27
- 239000000243 solution Substances 0.000 description 23
- 238000010828 elution Methods 0.000 description 18
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 12
- 238000005194 fractionation Methods 0.000 description 10
- 239000007864 aqueous solution Substances 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 7
- 230000000007 visual effect Effects 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 239000007853 buffer solution Substances 0.000 description 5
- 239000008363 phosphate buffer Substances 0.000 description 5
- 238000004364 calculation method Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 3
- 238000012442 analytical experiment Methods 0.000 description 2
- 238000004040 coloring Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000006978 adaptation Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000011982 device technology Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000000576 food coloring agent Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 229920002379 silicone rubber Polymers 0.000 description 1
- 229920003002 synthetic resin Polymers 0.000 description 1
- 239000000057 synthetic resin Substances 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
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- Sampling And Sample Adjustment (AREA)
Description
計測装置、分析装置の中のクロマトグラフに関する新規の装置開発。
クロマトグラフが数多く市販されているが、本発明はその中の分取を主目的とするフラクションコレクターに関する装置の発明である。Development of new equipment related to chromatography among measuring and analytical equipment.
There are many chromatographs available on the market, and the present invention is directed to a fraction collector, the main purpose of which is fraction collection.
精密用のクロマトグラフィーは専門メーカーにより数多く製品化され世界の研究機関に供給されている。近年はクロマトグラフの活用は精密分析のみならず、成分の分離精製にも及んでいる。その分離した成分のフラクションの方法として、新規のフラクションコレクター、技法はクロマトグラフィーに適した装置である。各研究機関では市販装置の利用がほとんどであり、自作したとしても、市販品のアレンジである。
現在のフラクションコレクター（市販品）は電動装置も含めたものであり、高価でもある。実験指導者や研究者には高価で購入する箇所は限られているのが現状である。それに、フラクションした成分の純度をさらに上げることは困難であるが、本装置技術ではフラクションした成分の純度を上げることが可能である。 分離メカニズムの特徴として溶離液の比重さをフラクション技法として活用していることである。
（なお、クロマトグラフは装置、クロマトグラフィーはその手法を全体、クロマトグラムは放出データーを記録したものを意味し分取装置をフラクションコレクターという）Many precision chromatography instruments are manufactured by specialized manufacturers and supplied to research institutes around the world. In recent years, the use of chromatography has expanded beyond precision analysis to include the separation and purification of components. As a method for fractionating the separated components, new fraction collectors and techniques are used that are suitable for chromatography. Most research institutes use commercially available instruments, and even if they make their own, they are simply adaptations of commercially available products.
Current fraction collectors (commercially available) include motorized devices and are expensive. They are expensive for lab instructors and researchers, and there are limited places where they can be purchased. In addition, it is difficult to further increase the purity of the fractionated components, but this device technology makes it possible to increase the purity of the fractionated components. A feature of the separation mechanism is that the specific gravity of the eluent is used as a fractionation technique.
(Note that chromatograph is the device, chromatography refers to the whole method, and chromatogram refers to the recording of emission data, and the fractionation device is called a fraction collector.)
従来のフラクションコレクターでは駆動のための電動式モーター類が必要であったが本装置、技術は分離溶液の比重差をのみを利用するためにその分離度が課題となった。
この分離度は液体装置付属の検出器とレコーダーにてクロマトグラムを得て判断できる。
そのクロマトグラムと今回発明の密度差の分離によるフラクションコレクター（ガラス管）の目視クロマトグラム（図１）とで比較する必要があった。
目視クロマトグラムをより明瞭にするために溶離液を緩衝液として濃度を変えて何回かにわたり、実験を行った。課題として次の事が上がった。
１．緩衝液の濃度を変えて分離度を上げて検出器とレコーダーにてクロマトグラム
と比較出来るようにする事。
２．成分分取の際、せっかく分離している成分が混ざり合わないようにする分取テクニックを取得する事。
３．より分離度を上げるためにグラジエント溶出法の溶離液の濃度影響調査。
４．従来のフラクションコレクターより純度を上げれる装置とするため、目視クロマトグラムを分離帯を三段階濃度に分取する必要がある。
５．従来のクロマトグラフィーのようにフラクションコレクターに比べて組立費用が格段に安いという特徴を証明するために、本装置を自作して費用を計算する事。Conventional fraction collectors require electric motors for drive, but this device and technology utilizes only the specific gravity difference of the separation solutions , so the degree of separation became an issue.
This degree of separation can be determined by obtaining a chromatogram using a detector and recorder attached to the liquid device.
It was necessary to compare this chromatogram with the visual chromatogram (Figure 1) of the fraction collector (glass tube) based on density difference separation of the present invention.
In order to make the visual chromatogram clearer, the experiment was carried out several times with different concentrations of buffer solution as the eluent. The following issues were raised:
1. By changing the concentration of the buffer solution, the degree of resolution can be increased and the chromatograms can be compared using a detector and recorder.
2. When separating components, acquire separation techniques that prevent the components that have been separated from each other from mixing together.
3. Investigating the effect of eluent concentration in gradient elution to further improve separation.
4. In order to make the fraction collector more pure than conventional fraction collectors, it is necessary to separate the visual chromatogram into three concentration levels.
5. To prove that the assembly cost is significantly lower than that of conventional chromatography using a fraction collector, build this device yourself and calculate the cost.
１．緩衝液の濃度を変えて分離度を上げて、検出器とレコーダーにてクロマトグラム
と比較出来るようにするための解決手段として、分取液の濃度変化時に成分ピークが現われるようにする事が大事で、その溶離液の濃度がある範囲で効果が認められるを実験した。
２．成分分取の際、せっかく分離している成分が混ざり合わないようにするテクニックを習得するために、分取技法を仕上げる。
３．より分離度を上げるためにグラジエント溶出法の活用が大切で２つ溶離液の濃度勾配を決める分析実験をした（ＵＶ検出器と目視ガラス管分離の比較をする）そして、従来のフラクションコレクターより純度を上げれる分取装置とするため、目視クロマトグラムを三段階濃度に分取する必要があり、その解決手段として、分離成分を分取し、上層、中層、下層に分け、ＨＰＬＣ－ＵＶ検出器で各層の濃度を測定する。
４．従来のフラクションコレクターよりも純度が上がった事を確認する計算法を作成するために本発明の分取用ガラス管内での分離帯を中央部を軸に上下同等の距離部を採取し、ＨＰＬＣ－ＵＶ検出器のクロマトグラムと比較計算をする。
５．従来のクロマトグラフィーのようにフラクションコレクターに比べて組立費用が格段に安いという特徴を証明するために、本装置の部品を調達し、弊社で自作して本装置の試作費用を計算した。分取用ガラス管は径を中小大と寸法を換えて試作してた。目視分離で再確認し、上述４、５の方法で判定した。1. In order to increase the degree of separation by changing the concentration of the buffer solution and to be able to compare the chromatograms with those of the detector and recorder, it is important to make the component peaks appear when the concentration of the separation solution changes. We conducted an experiment to see if this effect can be seen within a certain range of eluent concentrations.
2. When separating components, you will perfect your separation techniques to learn techniques to prevent the components that have already been separated from mixing together.
3. To improve the degree of separation, it is important to use gradient elution, and an analytical experiment was conducted to determine the concentration gradient of two eluents (comparing UV detector and visual glass tube separation). In order to create a fractionation device that can increase purity more than conventional fraction collectors, it is necessary to fractionate the visual chromatogram into three levels of concentration. As a solution to this, the separated components are fractionated and divided into upper, middle, and lower layers, and the concentration of each layer is measured with an HPLC-UV detector.
4. In order to create a calculation method to confirm that the purity is higher than that of conventional fraction collectors, samples are taken from the separation zone in the preparative glass tube of the present invention at equal distances above and below the center, and a comparison calculation is made with the chromatogram of the HPLC-UV detector.
5. In order to prove that the assembly cost is significantly lower than that of conventional chromatography using fraction collectors, we procured the parts for this device, built it ourselves, and calculated the cost of prototyping this device. The glass tubes for fractionation were prototyped with different diameters, small, medium, and large. They were reconfirmed by visual separation and judged using the methods 4 and 5 above.
１．溶離液に使用される濃度はリン酸緩衝液で０．０２０～０．０４０モルの範囲で分離が良い事を突き詰めた。この範囲を決める操作は通常の化学実験で行える簡単なものである。
２．フラクションされた部分について純度を上げるために必要部を抜き取れるために下部のガラスコックを細くして、滴下量を少なめにしていくと、分画液がほとんど混合されず分可能であった。この操作も複雑なものではない。
３．より分離度を上げるためにグラジエント溶出法の活用が大切で２つ溶離液の濃度勾配を決める分析実験をした結果、水溶性緩衝液と有機溶媒メタノールの混合比１２％～５８％の範囲で良好な結果を得た。緩衝液の濃度を上げ過ぎるとグラジエント溶出の際に沈殿を生じる。要因は溶解度に差があり過ぎるためである。今回は食品用の色素混合物で良き成果を得たが、他の成分例えばヌクレオチド類、ビタミン類でも溶離液を決めグラジエント溶出法を駆使すれば、同様な良き結果が得られることが推測される。（ＵＶ検出器のクロマトグラムと目視ガラス管比較をして判定出来る）
４．従来のフラクションコレクターより純度を上げれる分取装置とするため、目視クロマトグラムを三段階濃度に分取し実験上の計算法で判定した結果、本発明のフラクションコレクターはより純度が上がっていた。この成功によりより純度が必要な精密分析に試料作製に貢献する、例えばＨＰＬＣ－Ｍ ＡＳＳによる分子構造解析等。純度が上がった事を確認する計算法を作成するために本発明の分取用ガラス管内での分離。
帯を中央部を軸に上下同等の距離部を採取し、ＨＰＬＣ－ＵＶ検出器との比較計算をした結果、明らかにピーク中央部で純度上がっていた。
５．高価なフラクションコレクター装置を付属しなくても、従来のフラクションコレクターよりも分離度と精度が上がる事が証明出来たので、現状のフラクションコレクターは駆動装置を稼働させるために電気エネルギーを必要とするが本装置はガラスカラムあるいは合成樹脂カラムのみの接続で良い。
従来装置に比べて廉価なので、各種研究機関に格段に普及していくと考えられる。
現状段階で３２万円～３５万円程度の装置が本発明による装置では格段と安く概略１～２万円程度で仕上がる。 また従来のフラクションコレクターはフラクションの量を決めるために分取のタイムの種類やポンプ流量を変化させる必要があったが、新方式では分取ガラス管のサイズや長さを換えれば自由に幅広く分取できる。そして各装置に持ち運べる携帯用フラクションコレクターの開発になった。またより純度を上げる分取技法としてカラムにセプタムを付ける管も考えられる。1. We have found that the best separation results are achieved when the concentration of the eluent is in the range of 0.020 to 0.040 molar in phosphate buffer. The procedure for determining this range is simple and can be carried out in ordinary chemical experiments.
2. In order to increase the purity of the fractionated parts, the glass stopcock at the bottom was narrowed and the amount of dripping was reduced so that the necessary parts could be extracted, and the fractionated liquid was hardly mixed and could be separated. This operation is not complicated either.
3. To further improve the degree of separation, it is important to utilize gradient elution, and as a result of analytical experiments to determine the concentration gradient of two eluents, good results were obtained with a mixture ratio of water-soluble buffer solution and organic solvent methanol in the range of 12% to 58%. If the concentration of the buffer solution is increased too much, precipitation will occur during gradient elution. This is because there is too much difference in solubility. This time, good results were obtained with a food coloring mixture, but it is expected that similar good results can be obtained with other components, such as nucleotides and vitamins, by determining the eluent and making full use of the gradient elution method. (This can be determined by comparing the chromatogram from the UV detector with the glass tube with the naked eye.)
4. In order to make a fractionation device that can increase purity more than conventional fraction collectors, visual chromatograms were fractionated into three levels of concentration and judged by experimental calculation methods, and the fraction collector of the present invention had a higher purity. This success contributes to the preparation of samples for precision analysis that requires higher purity, such as molecular structure analysis by HPLC-MASS. Separation in the fractionation glass tube of the present invention was performed to create a calculation method to confirm the increase in purity.
Samples were taken from equal distances above and below the center of the band, and a comparison was made with the HPLC-UV detector. It was clear that the purity was higher in the center of the peak.
5. It has been proven that the separation and accuracy can be improved compared to conventional fraction collectors without the need for an expensive fraction collector. Current fraction collectors require electrical energy to operate the drive unit, but this device only requires the connection of a glass column or synthetic resin column.
Because it is less expensive than conventional devices, it is expected to become much more popular among various research institutions.
The equipment currently costs around 320,000 to 350,000 yen, but the equipment of the present invention is much cheaper, costing roughly 10,000 to 20,000 yen. In addition, while conventional fraction collectors require changing the type of fraction collection time and pump flow rate to determine the amount of fraction, the new method allows for a wide range of fraction collection by changing the size and length of the fraction collection glass tube. This led to the development of a portable fraction collector that can be carried to each device. Another fraction collection technique that can further increase purity is the addition of a septum to the column.
クロマトグラフは通常市販されている装置で良い、フラクションコレクター用の配管も通常の溶出液に使用する配管で十分である。
フラクションコレクター用のガラス管は汎用されているクロマト管でも良いが下部の付属フィルターは必要ないので、普通のガラス管下部にコックを付けて、流量が調節できる微細コックであればなお良い。上部にシリコンゴム栓をつけ、空気穴と配管穴を開けて使用するとなお良い。
具体的な図面、見取り図に示している器具を使用。いずれの器具も一般の化学実験室に備えられているもので特別なものではなく教育実験用として市販されている。
具体的な器具と操作について記す。
１、通常のクロマトグラフにグラジエント用の２溶離液（２種の溶離液は比重に違いのある溶離液を使用）をポンプに接続、検出器の出口にフラクションコレクターへの配管パイプを接続。そして実験を開始する。
２．ガラス管の下部コック閉めて、溶出液を順次、ガラス管に貯めてゆく、溶出が進むと成分が分離され分離層を生じる。分析終了後、送液ポンプをストップさせる。そして下部のコックを開けて順次、分離層を分取してゆく。
（分取作業は微量滴下するのが良い）
３．レコーダーのクロマトグラムと分取用ガラス管の分離層と比較し分離精度を確認する。The chromatography may be carried out by any commercially available device, and the piping for the fraction collector may be the same as that used for ordinary eluents.
The glass tube for the fraction collector can be a general-purpose chromatography tube, but since there is no need for an attached filter at the bottom, it is better to use a regular glass tube with a tap at the bottom and a fine tap that can adjust the flow rate. It is even better to use a silicon rubber stopper at the top and open an air hole and piping hole.
The equipment used is shown in the specific drawings and floor plans. All of the equipment is not special and is available commercially for educational purposes, as it is found in most chemistry laboratories.
Specific equipment and procedures are described.
1. Connect two gradient eluents (two eluents with different specific gravities) to a pump in a normal chromatograph, and connect a pipe to a fraction collector at the detector outlet. Then start the experiment.
2. Close the bottom cock of the glass tube and gradually collect the eluate in the glass tube. As the elution proceeds, the components separate and separate layers are formed. After the analysis is completed, stop the liquid delivery pump. Then open the bottom cock and gradually collect the separated layers.
(It is best to use minute drops for fractionation.)
3. Check the separation accuracy by comparing the chromatogram on the recorder with the separation layer in the preparative glass tube.
食品添加物用色素混合物の分取
「実験条件」
（１液）２０ｍｍｏｌリン酸緩衝液，５％メタノール水溶液、ｐＨ６．２５
（２液）１％酢酸，９５％メタノール水溶液
リニアグラジェント溶出法１液から２液まで２４分間とする
流速１．０ｍｌ
カラムＯＤＳ系カラム
（器具・装置）
通常の液体クロマトグラフと分取ガラス管
（内径１０ｍｍ，全長２００ｍｍ）
「試料」
食添色素５種を使用
５つの色素の分離度（Ｒｔ）の表を作成した。
分離帯においてほとんど分離どの向上がみられる。図３。Experimental conditions for separating food additive color mixture
(Solution 1) 20 mmol phosphate buffer, 5% methanol aqueous solution, pH 6.25
(2nd solution) 1% acetic acid, 95% methanol aqueous solution Linear gradient elution method 24 minutes from 1st solution to 2nd solution Flow rate 1.0 ml
Column ODS column (instrument/equipment)
Conventional liquid chromatograph and preparative glass tubes
(inner diameter 10 mm, total length 200 mm)
"sample"
Five types of food colorings were used. A table of the separation degree (Rt) of the five colorings was created.
Most of the improvement in separation is seen in the separation zone.
リニアグラジエントの差による分離度の差
「実験条件」
（１液）２０ｍｍｏｌリン酸緩衝液，５％メタノール水溶液、ｐＨ６．２５
（２液）１％酢酸，９５％メタノール水溶液
グラジェント溶出法Ａ法１液５％→１液８０％（２５分）
グラジェント溶出法Ｂ法１液１２％→１液９５％（２５分）
流速１．０ｍｌ
カラムＯＤＳ系カラム
（器具・装置）
通常の液体クロマトグラフと分取ガラス管（内径１０ｍｍ，全長２００ｍｍ）
(Solution 1) 20 mmol phosphate buffer, 5% methanol aqueous solution, pH 6.25
(2nd solution) 1% acetic acid, 95% methanol aqueous solution Gradient elution method A 1st solution 5% → 1st solution 80% (25 minutes)
Gradient elution method B: 12% solution 1 → 95% solution 1 (25 min)
Flow rate 1.0 ml
Column ODS column (instrument/equipment)
A conventional liquid chromatograph and a glass tube for fractionation (inner diameter 10 mm, total length 200 mm)
緩衝液濃度差による分離度
「実験条件」
１、低濃度
（１液）１５ｍｍｏｌリン酸緩衝液，５％メタノール水溶液、ｐＨ６．２５
（２液）１％酢酸，９５％メタノール水溶液
グラジェント溶出法Ａ法１液５％→１液８０％（２５分）
グラジェント溶出法Ｂ法１液１２％→１液９５％（２５分）
２、高濃度
（１液）４０ｍｍｏｌリン酸緩衝液，５％メタノール水溶液、ｐＨ６．２５
（２液）１％酢酸，９５％メタノール水溶液グラジェント溶出法Ａ法
１液５％→１液８０％（２５分）グラジェント溶出法
流速１．０ｍｌカラムＯＤＳ系カラム
（器具・装置）
通常の液体クロマトグラフと分取ガラス管（内径１０ｍｍ，全長２００ｍｍ）
濃度勾配に差があり濃度も高い方が分離騒帯は分離向上している。本実験の技法には適していることを示している。ＢとＣ、ＣとＤで大きな差がある。
他の成分についてもグラジェント溶出法の溶離液設定は大切で溶出条件の適したものを選ぶことにより分離度の良き層を得ることが出来る。図５。Separation degree due to difference in buffer concentration "Experimental conditions"
1. Low concentration (1st solution) 15 mmol phosphate buffer, 5% methanol aqueous solution, pH 6.25
(2nd solution) 1% acetic acid, 95% methanol aqueous solution Gradient elution method A 1st solution 5% → 1st solution 80% (25 minutes)
Gradient elution method B: 12% solution 1 → 95% solution 1 (25 min)
2. High concentration (1st solution) 40 mmol phosphate buffer, 5% methanol aqueous solution, pH 6.25
(2nd solution) 1% acetic acid, 95% methanol aqueous solution Gradient elution method A method 1st solution 5% → 1st solution 80% (25 min) Gradient elution method Flow rate 1.0 ml Column ODS column (instrument/equipment)
A conventional liquid chromatograph and a preparative glass tube (inner diameter 10 mm, total length 200 mm)
The higher the concentration gradient, the better the separation of the separation zones. This shows that this technique is suitable for this experiment. There is a big difference between B and C, and between C and D.
For other components, the eluent settings for gradient elution are also important, and by selecting appropriate elution conditions, layers with good separation can be obtained.
液体クロマトグラフィーは科学研究には欠かせない重要な分析装置であり、世界の研究機関で活用されている。この装置の中で本発明のフラクションコレクターは有用な成分を混合物から取り出す技術として必要不可欠のものである。
有用な成分は分取後、次の研究に供される。例えば、生理活性測定や分子構造解析などへ。
図面でも示しましたが、従来の装置は電動式で価格も高い、今回の装置は価格が格段と安く、世界の各研究所に普及すると考えられる。その上に溶離液の密度差による分離なので、再分取可能で、従来装置では出来なかったより精密分取も可能にした。ゆえに本発明の溶解後式フラクションコレクターは、すぐにでも産業上有効に利用出来るものである。本装置、技術は精密分析や有用物質の分離精製に適している。
そして、各装置に持ち運べる携帯用フラクションコレクターとして使用出来。
ガラス管にセプタムを装着してより精度よく抜き取ることも出来、より精密分析法に有効である。図６。Liquid chromatography is an important analytical tool indispensable for scientific research and is used in research institutions around the world. The fraction collector of the present invention is an essential tool for extracting useful components from a mixture.
After separating the useful components, they are used for further research, such as bioactivity measurements and molecular structure analysis.
As shown in the drawing, conventional devices are electric and expensive, but the cost of this device is significantly cheaper and it is expected that it will become widespread in laboratories around the world. Furthermore, because separation is based on the density difference of the eluent, it is possible to re-fraction and to perform more precise fractionation than was possible with conventional devices. Therefore, the post-dissolution type fraction collector of this invention can be effectively used in industry immediately. This device and technology are suitable for precision analysis and the separation and purification of useful substances.
It can also be used as a portable fraction collector that can be carried to each device.
By attaching a septum to the glass tube, it is possible to extract the sample more accurately, which is effective for more precise analysis.
Ａ ・・・ 試料の分離帯
Ｂ ・・・ 試料の分離帯
Ｃ ・・・ 試料の分離帯
Ｄ ・・・ 試料の分離帯
Ｅ ・・・ 試料の分離帯
Ｓ ・・・ スタート
（１）・・・ シリンジ用セプタｍｕA... Sample separation band B... Sample separation band C... Sample separation band D... Sample separation band E... Sample separation band S... Start (1)... Syringe septum mu
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